JPH0441499A - Carcinogenesis-preventing agent - Google Patents

Abstract

NEW MATERIAL:2-Hydroxy-3-methoxy-5-(2-propenyl)phenol 1-0-beta-D-(6' -galloyl) glucopyranoside expressed by the formula. USE:The compound expressed by the formula can be utilized in order to prevent carcinogenesis as an anti-carcinogeness promoter, since the compound strongly suppresses Epstein-Barr virus activity by tetradecanoylphorbol acetate. PREPARATION:Allspice is extracted with methanol and the resultant extract is distributed into chloroform and water. The resultant water layer is distributed using acetic acid ethyl ester and water and the acetic acid ethyl ester layer is repeatedly separated and purified with a normal-phase or reverse-phase column chromatography to provide the compound expressed by the formula.

Description

DETAILED DESCRIPTION OF THE INVENTION (Industrial Field of Application) The present invention relates to a carcinogenesis preventive agent, and more particularly to a novel carcinogenesis preventive agent having an anti-carcinogenic promotion effect.

(Problems to be solved by the prior art and the invention) It has been pointed out that early detection and early treatment of adult diseases and cancer are necessary, but most adults are exposed to various substances including carcinogenic chemicals. It is thought that normal cells have already been damaged by carcinogenic factors, and as a result, normal cells harbor potential tumor cells that have changed almost irreversibly. It is also generally accepted that these latent tumor cells undergo further double acid effects and transform into tumor cells, and this process is called promotion.

On the other hand, Epstein-Barr virus (EBV), which belongs to the herpesvirus family, is known as a virus that is said to cause Burkitt's lymphoma and nasopharyngeal cancer. The virus is common only to these cancer patients,
It is said that almost all adults are infected with EBV. EBV has been shown to infect normal human B lymphocytes, turn them into blast cells, and endow them with unlimited proliferation potential, and is a resident resident of humans that has built-in tumorigenicity to B lymphocytes. Defined as a viral factor. EBV-infected B lymphocytes undergo the promotion described above and migrate to cancer cells, and epidemiological experiments have shown that phorbol esters such as tetradecanoylphorbol acetate (TPA) are present as promoters. It has also been proven. For example, plants in the Euphorbiaceae family that are distributed in East Africa contain phorbol esters, and the surrounding soil, crops, and drinking water are contaminated with the phorbol esters secreted by these plants and are constantly exposed to these. Burkitt's lymphoma is occurring frequently in the area. Therefore, it is extremely important how to prevent such promotion in daily life, and the importance of primary child prevention centered on improving dietary habits has recently been particularly emphasized. The present inventors have focused on the promotion process of chemical carcinogenesis from the viewpoint of carcinogenesis prevention, and have been searching for substances that suppress carcinogenesis promotion from natural resources. It has been found that aldehydes such as polygodial and monoglycerides such as monolyruine from pearl barley can serve as anti-carcinogenic promoters. However, the search for naturally occurring anti-carcinogenic promoters has only just begun, and it cannot be said that sufficient studies have been conducted even on plants that are commonly used in daily life. It has been desired to provide an effective carcinogenic preventive agent.

(Means for Solving the Problems) The present inventors have developed a system using Raji (Raji), which is a Burkitt's lymphoma-derived 8128 cell culture containing the EBV genome.
) strain and used an experimental system with TPA as the oncogenic promoter to search for naturally occurring anti-carcinogenic promoters. More specifically, we searched for phase promoters in the expression of TPA and promotion activity in the Raji strain culture system. More specifically, the Raji strain culture system is cultured with n-butyric acid, which has a synergistic effect on the expression of promotion activity with TPA, and a test substance, and the EB virus early antigen, which is activated by TPA and expressed on the cell surface, is cultured. (EBV-EA) is observed by indirect fluorescent antibody method using antibodies derived from nasopharyngeal cancer patient serum. Most of the EBV activation inhibitors discovered by the authors suppress tumor development even in two-step carcinogenesis experiments using TPA, and are effective as anti-carcinogenic promoters (Kuroyo, et al., author: Antimutagenesis).・And Anticarcinogenesis Mechanisms ■, Blenheim Press, pp. 425-429, 1990
Year).

As a result of intensive research, the present inventors found that 2-hydroxy-
3-methoxy-5-(2-propenyl)phenol 1
-0-β-D-(6'-galoyl)glucopyranoside (2-hydroxy-3-methoxy-5-(2
-propenyl)phenol 1-0-β-D-
(6'-galloyl)glucopyranos
idc) [Structural Formula I] was discovered as a new compound, and the problem was solved by discovering a cancer prevention agent containing the compound as an active ingredient.

The compound is derived from Pimenta officinalis or Pimenta dioica of the family Myrtaceae.
), and especially in allspice whose seeds are dried (also known as pimento, vimenta, and jamaica vetuba). Extraction production from these plants can be achieved by a combination of known solvent extraction methods and various chromatography methods. For example, the extract obtained by extracting allspice with methanol is partitioned between chloroform and water. The resulting aqueous layer is then partitioned between ethyl acetate and water, and the ethyl acetate layer is repeatedly separated and purified by normal-phase and reverse-phase column chromatography to obtain the compound. Plant extracts containing the compound as an active ingredient can also be used as cancer prevention agents. It can be obtained by extraction using an organic solvent such as thylethyl ketone or a mixed solvent thereof. Furthermore, if necessary, it is also possible to obtain both components in which the active ingredient is concentrated by known separation means.

The compound thus obtained or the plant extract containing the compound can be used as a cancer prevention agent in the form of a composition containing an effective and non-toxic amount thereof. For example, oral preparations can be used as solid preparations such as tablets, capsules, troches, granules, and powders, or liquid preparations such as solutions and syrups. The proportion of the compound in the carcinogenic preventive agent of the present invention varies depending on the dosage form, but when ingested orally, the appropriate proportion is usually 0.3 to 15% by weight. The intake amount varies depending on the desired preventive effect and age, but for adults, the above-mentioned compound is usually used in the range of 0.5 to 5000 mg per day.

Next, the method for producing the compound of the present invention and its EBV activation inhibitory effect will be explained using test methods and examples.

(Test method) RPM11640 plus fetal serum and antibiotics was used as a culture medium for EBV latently infected human litoriblastoid cell line Raji cells. Under this culture condition, EBV-
The spontaneous EA expression rate is 0.1% or less. IX1
Raji cells adjusted to a concentration of 0' cells/ml were added to 4 m
The cells were cultured at 37° C. for 48 hours in the above culture solution containing M n-butyric acid, 20 ng/ml TPA, and the test substance at a concentration of 1 to 100 μg/ml. EBV-EA was stained by indirect fluorescent antibody method using nasopharyngeal cancer patient serum, and the percentage of positive cells was calculated relative to a control to which no test substance was added, and was taken as EBV activation inhibitory activity.

(Example 1) Allspice (1950g) was soaked in methanol,
After a week, this was filtered, and the solvent was distilled off from the filtrate by concentration to obtain an extract (157 g). Using this, the EBV activation inhibitory activity was measured according to the above test method (Table 1) (Example 2) The extract (157 g) obtained in Example 1 was partitioned between water and n-hexane, and the hexane layer was separated from the hexane layer. The solvent is distilled off and the extract (
57g) was obtained. Using this, EB according to the above test method
V activation suppression activity was measured (Table 1).

(Example 3) The extract obtained by distilling off the solvent of the aqueous layer obtained in Example 2 was distributed between chloroform and water, and the solvent was distilled off from the chloroform layer to obtain an extract (2-6 g). . Using this, EBV activation inhibitory activity was measured according to the above test method (
Table 1).

(Example 4) The extract obtained by distilling off the solvent of the aqueous layer obtained in Example 3 was distributed between ethyl acetate and water, and the solvent was distilled off from the ethyl acetate layer to obtain an extract ( 16-7g)
I got it. Using this, EBV activation inhibitory activity was measured according to the above test method (Table 1) (Example 5) The extract obtained in Example 3 (16-7 g) was subjected to silica gel column chromatography, and chloroform/methanol /water = 65: 35: 10 (V/V) The fraction fractionated with the lower layer solvent system was roughly purified by reverse phase liquid chromatography to obtain water/methanol =
A single compound was obtained from the fraction eluted with a 2:1 (V/V) solvent.

This compound is 2-hydroxy-3-methoxy-5-(2-propenyl)phenol 1-0-β-D-( It was identified as 6' galloyl) glucopyranoside.

FAB-MS (m/z): 495 (senior + 1),
315.307.297.289.180.153.1
36.107.89°IR (NaCL, cm-”): 3
425.1700.1620.1520.1460.1
325.1240.1080゜” H-NMR (CDC
I, ): 63.04 (211, J-6-811z, 7
), 3-30-3-35 (3I1.v, 2°, 3'
, 4'), 3-65 (III, dd, J=6nz,
6Hz, 5″), 3.71 (311,s, 3)
, 4-30 (III, dd, J=6nz, 11
11z, 6°a), 4-46 (1r1.d, J=11
11z, 6°b).

C67 (III, d-1ike, J=6.811z
, 1'), 4.88 (III, s, 9tra
ns), 4.93(Ill, 4. J=7IIz,
9-cis), 5.85 (III, vx, 8),
6-45 (III, d, J=2I[z, 4),
6.46 (III, s, 6), 6-99 (21
1, s, 2", 6").

"C-NMR (CDC1,): δ 40 (7),
55-8 (OMe), 63.4 (6”), 69
．． 8 (4'), 73.2 (2°), 73.9 (5
°).

75-4 (3°), 102-4 (1”), 107-
3 (4), 108.5 (2", 6" included 109-3 (6
), 115.1 (9), 119-2 (4”), 129-
8(5).

134, 3(2), 13'l 7(8), 138.
5 (1”), 145.4 (1).

145.5 (3", 5°), 14'l 8 (3),
165-6 (CO).

The attribution of these NMR signals is! )1 1 H and IH13C two-dimensional NMR spectra and remote 1H1
Confirmed by 3c two-dimensional NMR spectrum.

Using this, EBV activation inhibitory activity was measured according to the above test method.

Table 1 EBV activation inhibitory activity of test substance Test substance (8 g) Extract of Example 1 Extract of Example 2 Extract of Example 3 Extract of Example 4 Extract of Example 5 Compound (Effect of the invention) Shown in Table 1 As such, the 2-hydroxy-3-methoxy-5-(2-propenyl)phenol of the present invention 1-0-
β~D-(6'-galoyl)glucopyranoside is TPA
Since it strongly suppresses EBV activation by , it can be expected to be used as an anti-carcinogenic promoter for the purpose of preventing cancer development, and can also be used in cancer prevention drugs and cancer prevention foods.

Claims (2)

[Claims] (1) A glycoside represented by the structural formula I below. ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ Structural formula I (2) A carcinogenic preventive agent containing the glycoside according to claim 1 as an active ingredient.